Tax interacts with the cellular CREB protein and facilitates the binding of the coactivator CBP, forming a multimeric complex on the CRE-like sites in the HTLV-I promoter. The trimeric complex is believed to recruit additional regulatory proteins to the HTLV-I LTR, but there has been no direct evidence that CBP is required for Tax-mediated transactivation. We present evidence that Tax and CBP activate transcription from the HTLV-I 21 bp repeats on naked DNA templates. Transcriptional activation of the HTLV-1 sequences required both Tax and CBP and could be mediated by either the N-terminal activation domain of CBP or the full length protein. Fluorescence polarization binding assays indicated that CBP does not markedly enhance the affinity of Tax for the trimeric complex. Transcription analyses suggest that CBP activates Tax-dependent transcription by promoting transcriptional initiation and reinitiation. The ability of CBP to activate the HTLV-1 promoter does not involve the stabilization of Tax binding, but rather depends upon gene activation properties of the co- activator that function in the context of a naked DNA template.Recent studies have shown that the p300/CBP binding protein-associated factor (PCAF) is involved in transcriptional activation. PCAF activity has been shown strongly associated with histone acetyltransferase (HAT) activity. Intriguing new data from this laboratory suggest the presence of a HAT-independent transcription function that is activated in the presence of the HTLV-I Tax protein. In vitro and in vivo GST-Tax pull down and co-immunoprecipitation experiments demonstrate that there is a direct interaction between Tax and PCAF, independent of p300/CBP. PCAF can be recruited to the HTLV-I TRE site in the presence of Tax and PCAF cooperates with Tax in vivo to activate transcription from the HTLV-I LTR over 10-fold. Point mutations at Tax amino acid 318 (TaxS318A) or 319-320 (Tax M47) which have decreased or no activity on HTLV-I promoter, are defective for PCAF binding. Strikingly, the ability of PCAF to stimulate Tax transactivation is not solely dependent upon the PCAF HAT domain. Two independent PCAF HAT mutants, which knock out acetyltransferase enzyme activity, activate Tax transactivation to approximately the same level as wild-type PCAF. In contrast, p300 stimulation of Tax transactivation is HAT-dependent. These studies provide experimental evidence that PCAF contains a coactivator transcription function independent of the HAT activity on the viral LTR. - Gene regulation, Leukemia, Oncogenes, Retroviruses, Transcription Factors, Virus-Cell Interactions,

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Intramural Research (Z01)
Project #
1Z01BC005254-18
Application #
6289081
Study Section
Special Emphasis Panel (LRBG)
Project Start
Project End
Budget Start
Budget End
Support Year
18
Fiscal Year
1999
Total Cost
Indirect Cost
Name
National Cancer Institute Division of Basic Sciences
Department
Type
DUNS #
City
State
Country
United States
Zip Code
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